Project Summary In spite of the prevalence and severity of many neurological disorders, the development of new classes of drugs has been sluggish for 50 years. This is due largely to the lack of good model systems and tools to rapidly study relevant electrical and synaptic phenotypes. We aim to overcome these challenges. Recent advances in induced pluripotent stem cell (iPSC) technology reveal the first prospects for studying human neurons paired with clinical histories using fast in vitro technologies. The complex electrophysiological behavior of these cell can be recorded with the Optopatch platform and microscope systems recently developed at Q-State. With these tools, it is possible to simultaneously stimulate (blue light) and record (red light) electrical activity from around a hundred neurons with one millisecond temporal resolution, single cell spatial resolution, and high signal to noise. This system can be used to measure single cell excitability and firing patters or to probe synaptic transmission by stimulating a subset of neurons with spatially patterned blue light. Moving forward, we propose to increase microscope throughput without sacrificing capabilities and rigorously test the platform?s performance. First, the microscope will be upgraded with advanced environmental controls, 96- well plate compatibility, and a fluid-handling robot for compound addition. Next, data storage and analysis will be securely moved to the cloud to handle 2.5 Terabyte/day data rates. Once the microscope is fully functional, sensitivity and reproducibility (well to well, plate to plate, and batch to batch) will be tested using a library of control compounds. Finally, as a first application, we will search for a robust, screenable phenotype for Dravet syndrome. Neurons will be prepared from ten healthy and ten Dravet patients to look for differences in firing that transcend variation in the genetic background. A drug that ameliorates the disease phenotype in the majority of cell lines is a promising candidate to be broadly effective in the clinic. A well validated, high-throughput electrophysiology platform with confirmed phenotypic readouts in human iPSC disease neurons has the potential to change the drug screening landscape for neurological disorders. We hope to open a new path to finding treatments for these horrible diseases.